Clone and Sequence Analysis on cDNA of Ghrelin Gene from Guangxi Buffalo

[Objective] The study was to clone cDNA of ghrelin gene from Guangxi buffalo and analyze its sequence.[Method]With the total RNA from true stomach tissue of Guangxi swamp buffalo as the template,the specific primers were taken to amplify cDNA of ghrelin gene.The amplified products were connected with pMD 18-T and transformed into Escherichia coli DH5α after it was recovered and purified with gel.The positive clones were screened out from transformation products after they were identified by PCR and double digestion.The positive clones were taken to do sequence analysis after they were cultured by liquid medium of 5×1 ml LB.[Result]The cDNA of ghrelin gene from Guangxi buffalo was successfuly gained in the test,and the length of cDNA fragment was 339 bp and it coded 113 amino acid.The result of BLAST analysis showed that the fragment only had 6 basyl difference with that of prepro ghrelin gene from yellow cattle and their homology was 98.2%.The ghrelin gene of Guangxi buffalo and yellow cattle were all composed by 27 amino acid,and the sequence homology of their ghrelin genes was 100%.[Conclusion] The test provided the theoretical basis for further study on biological function of ghrelin gene.